The detection, identification and quantification of nucleic acids is central to a number of technologies having application in diverse industries including the medical, agricultural, pharmaceutical, biotechnological, and security fields.
Real time polymerase chain reaction (RT-PCR) is a powerful technology for such detection, identification, and quantification of nucleic acids. Traditional RT-PCR methods measure total DNA using fluorescent intercalation dyes or labelled primers. Electrochemical measurement in principle offers many advantages, in instrumentation, measurement system design and cost of implementation. There have been a number of reports of coupling of electrochemical measurement with PCR amplification. These methods, again, have either used intercalation reagents for non-specific measurement or electrochemical labels on the primers or nucleotides. The sensitivity (signal/noise or signal/background) of the measurement technique determines the number of cycles of amplification required to obtain a reliable signal, which in turn determines the time to result and also the influence of replication errors.
There remains a need for methods of detecting amplified nucleic acid at high sensitivity, and particularly methods that are robust and reliable and/or amenable to implementation in high throughput or microfluidic systems.
The present invention is directed to providing methods and apparatuses for amplifying, detecting and/or quantifying nucleic acids, or to at least provide the public with a useful choice.